Enhanced resistance to Botrytis cinerea in genetically-modified Vitis vinifera L. plants over-expressing the grapevine stilbene synthase gene

Abstract

Grapevine (Vitis vinifera L.) was genetically modified with a construct containing a cDNA insert encoding the stilbene synthase gene (Vst1) from grapevine, under the control of the cauliflower mosaic virus 35S promoter in order to test the potential of over-production of resveratrol to protect plants from fungal attack. Southern blot hybridization and quantitative real-time PCR analysis demonstrated the presence and integration of one copy of exogenous DNA sequences in two grapevine-modified lines. Relative expression of the Vst1 gene in different modified lines was confirmed by using gene-specific quantitative real-time PCR. Compared to the control, the concentration of trans-resveratrol quantified by HPLC was up to 7.5 fold higher in the modified plants. The necrotic lesion size of leaves of intact modified plants inoculated by Botrytis cinerea B05.10 strain was consistently smaller and significantly different (p B 0.05) than in control plants, showing that modified grapevine plants were more resistant to the pathogen than the control plants.

Grapevine Grafting: Scion Transcript Profiling and Defense-Related Metabolites Induced by Rootstocks

“…Stilbenoids show ROS scavenging activity acting as a powerful defense system against biotic stresses (e.g., powdery mildew, downy mildew; Dabauza et al, 2015 ; Houillé et al, 2015 ).…”

“…The authors showed variations in modulation of genes involved in the phenylpropanoid pathway (mainly stilbene and flavonoid genes), suggesting a resveratrol-mediated way to enhance water stress resilience in a tolerant rootstock genotype. Stilbenoids (resveratrol and viniferin in particular) have reactive oxygen species (ROS) scavenging activity and act as a powerful defense system against biotic stresses (e.g., powdery mildew, downy mildew; Dabauza et al, 2015 ; Houillé et al, 2015 ). Furthermore, it is well-known that Vitis vinifera represents the best nutritional source of stilbenoid compounds with valuable effects on human health; these compounds have numerous pharmacological activities including anti-fungal, anti-inflammatory and anti-tumor effects (Bai et al, 2010 ; Zhang et al, 2010 ; Houillé et al, 2014 ).…”

Agrobacterium‐mediated transformation of Vitis Cv. Monastrell suspension‐cultured cells: Determination of critical parameters

“…The npt II gene confers resistance to aminoglycoside antibiotics such as kanamycin and paromomycin. In most studies on transformation of grapevine SCC kanamycin has been the most frequently used selection agent although at very diverse concentrations from 10 to 100 mg L −1 , probably depending on the Vitis genotype. In the present study we found that Monastrell cells are almost insensitive to kanamycin and the selection was carried out with paromomycin, since this antibiotic had been previously used by other authors to select transformed grapevine tissues .…”

Biotechnological Approaches: Gene Overexpression, Gene Silencing, and Genome Editing to Control Fungal and Oomycete Diseases in Grapevine

“…In vitro infection tests with Botrytis cinerea were carried out on grapevine micro cuttings transformed with the V. vinifera Vst1 gene under the control of a promoter inducible by fungal infection (PR10 promoter isolated from alfalfa); the results obtained confirmed an enhanced resistance against the pathogen in the transgenic lines that exhibited the highest resveratrol synthesis level [ 51 ]. In another study, the expression of the Vst1 gene under the cauliflower mosaic virus 35S promoter resulted in smaller botrytis necrotic lesion size in transgenic grapevine in comparison with control plants, demonstrating the existence of a positive correlation among resveratrol content and resistance to Botrytis cinerea [ 52 ]. In transgenic Arabidopsis plants, the expression of the Vitis pseudoreticulata STS gene ( VpSTS ), encoding a novel stilbene synthase, led to greater resistance to powdery mildew, since its expression seemed to be stimulated by SA, as happens with the expression of its homologous allele from susceptible Vitis vinifera [ 56 ].…”